Optimization and Validation of Amine Coupling of Ligand

In this chapter, we focus on the ideal strategy to optimize and validate amine coupling for SPR studies. In particular, we provide an in-depth description of each and every component required to establish validation before the screening of fragments. The primary emphasis is to maintain the protein in its natural state without losing its conformational change. The majority of proteins contain multiple amine groups so efficient immobilization of the protein is usually easily achieved without denaturation and inactivation of the protein. Optimization and validation protocol is presented here with important criteria and troubleshooting with action taken considered at each step.